Dissociation of pathological and molecular phenotype of variant Creutzfeldt-Jakob disease in transgenic human prion protein 129 heterozygous mice (original) (raw)
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Acta Neuropathologica Communications, 2016
Introduction: Mammalian prions are proteinaceous pathogens responsible for a broad range of fatal neurodegenerative diseases in humans and animals. These diseases can occur spontaneously, such as Creutzfeldt-Jakob disease (CJD) in humans, or be acquired or inherited. Prions are primarily formed of macromolecular assemblies of the disease-associated prion protein PrP Sc , a misfolded isoform of the host-encoded prion protein PrP C. Within defined host-species, prions can exist as conformational variants or strains. Based on both the M/V polymorphism at codon 129 of PrP and the electrophoretic signature of PrP Sc in the brain, sporadic CJD is classified in different subtypes, which may encode different strains. A transmission barrier, the mechanism of which remains unknown, limits prion cross-species propagation. To adapt to the new host, prions have the capacity to 'mutate' conformationally, leading to the emergence of a variant with new biological properties. Here, we transmitted experimentally one rare subtype of human CJD, designated cortical MM2 (129 MM with type 2 PrP Sc), to transgenic mice overexpressing either human or the VRQ allele of ovine PrP C. Results: In marked contrast with the reported absence of transmission to knock-in mice expressing physiological levels of human PrP, this subtype transmitted faithfully to mice overexpressing human PrP, and exhibited unique strain features. Onto the ovine PrP sequence, the cortical MM2 subtype abruptly evolved on second passage, thereby allowing emergence of a pair of strain variants with distinct PrP Sc biochemical characteristics and differing tropism for the central and lymphoid tissues. These two strain components exhibited remarkably distinct replicative properties in cell-free amplification assay, allowing the 'physical' cloning of the minor, lymphotropic component, and subsequent isolation in ovine PrP mice and RK13 cells. Conclusions: Here, we provide in-depth assessment of the transmissibility and evolution of one rare subtype of sporadic CJD upon homologous and heterologous transmission. The notion that the environment or matrix where replication is occurring is key to the selection and preferential amplification of prion substrain components raises new questions on the determinants of prion replication within and between species. These data also further interrogate on the interplay between animal and human prions.
Prion Strain Characterization of a Novel Subtype of Creutzfeldt-Jakob Disease
Journal of virology, 2017
In 2007, we reported a patient with an atypical form of Creutzfeldt-Jakob disease (CJD) heterozygous for methionine-valine (MV) at codon 129 who showed a novel pathological prion protein (PrP(TSE)) conformation with an atypical glycoform (AG) profile and intraneuronal PrP deposition. In the present study, we further characterize the conformational properties of this pathological prion protein (PrP(TSE) MV(AG)), showing that PrP(TSE) MV(AG) is composed of multiple conformers with biochemical properties distinct from those of PrP(TSE) type 1 and type 2 of MV sporadic CJD (sCJD). Experimental transmission of CJD-MV(AG) to bank voles and gene-targeted transgenic mice carrying the human prion protein gene (TgHu mice) showed unique transmission rates, survival times, neuropathological changes, PrP(TSE) deposition patterns, and PrP(TSE) glycotypes that are distinct from those of sCJD-MV1 and sCJD-MV2. These biochemical and experimental data suggest the presence of a novel prion strain in C...
Acta Neuropathologica Communications
The methionine (M)—valine (V) polymorphic codon 129 of the prion protein gene (PRNP) plays a central role in both susceptibility and phenotypic expression of sporadic Creutzfeldt-Jakob diseases (sCJD). Experimental transmissions of sCJD in humanized transgenic mice led to the isolation of five prion strains, named M1, M2C, M2T, V2, and V1, based on two major conformations of the pathological prion protein (PrPSc, type 1 and type 2), and the codon 129 genotype determining susceptibility and propagation efficiency. While the most frequent sCJD strains have been described in codon 129 homozygosis (MM1, MM2C, VV2) and heterozygosis (MV1, MV2K, and MV2C), the V1 strain has only been found in patients carrying VV. We identified six sCJD cases, 4 in Catalonia and 2 in Italy, carrying MV at PRNP codon 129 in combination with PrPSc type 1 and a new clinical and neuropathological profile reminiscent of the VV1 sCJD subtype rather than typical MM1/MV1. All patients had a relatively long durati...
Modulation of Creutzfeldt-Jakob disease prion propagation by the A224V mutation
Annals of neurology, 2015
Mutations in the gene encoding the prion protein (PrP) are responsible for approximately 10-15% of cases of prion disease in humans, including Creutzfeldt-Jakob disease (CJD). Here we report the discovery of a previously unreported C-terminal PrP mutation (A224V) in a CJD patient exhibiting a disease similar to the rare VV1 subtype of sporadic CJD and investigate the role of this mutation in prion replication and transmission. We generated transgenic (Tg) mice expressing human PrP with the V129 polymorphism and A224V mutation, denoted Tg(HuPrP,V129,A224V) mice, and inoculated them with different subtypes of sporadic (s) CJD prions. Transmission of sCJD VV2 or MV2 prions was accelerated in Tg(HuPrP,V129,A224V) mice compared to Tg(HuPrP,V129) mice, with incubation periods of ∼110 days and ∼210 days, respectively. In contrast, sCJD MM1 prions resulted in longer incubation periods in Tg(HuPrP,V129,A224V) mice compared to Tg(HuPrP,V129) mice (∼320 days v. ∼210 days). Prion strain fidelit...
No Adaptation of the Prion Strain in a Heterozygous Case of Variant Creutzfeldt-Jakob Disease
Emerging Infectious Diseases, 2020
I n 2016, a definite case of clinical variant Creutzfeldt-Jakob disease (vCJD) in a person heterozygous for methionine/valine (MV) at codon 129 of the prion protein gene (PRNP 129MV) was reported in the United Kingdom (1). Given the relatively atypical clinical features in this case, we considered it important to ascertain the strain of prion agent to determine whether there had been strain adaption or whether the patient's genetic background may have influenced the disease phenotype. We conducted a study to determine whether we could isolate the same prion strain from this case of vCJD in a 129MV individual as was identified in previous 129 methionine homozygous (129MM) genotype vCJD cases, consistent with the hypothesis of a causal link to bovine spongiform encephalopathy (BSE). The clinical features for this patient were consistent with a diagnosis of either vCJD or sporadic Creutzfeldt-Jakob disease (sCJD). Results from magnetic resonance imaging (MRI) of the patient's brain were suggestive of sCJD on diffusion-weighted imaging (DWI) sequences, although the single coronal fluid-attenuated inversion recovery (FLAIR) sequence in this case was not diagnostic because of movement artifact. Results of cerebrospinal fluid (CSF) real-time quaking-induced conversion assay analysis and the direct detection assay for vCJD infection in the blood were negative. However, at autopsy, neuropathological examination revealed florid plaques, and biochemical analysis of prion protein (PrP) from the brain confirmed a type 2B profile, both characteristic of vCJD (1). Abnormal PrP was also detected in peripheral tissues. Recent studies in which researchers used protein misfolding cyclic amplification in CSF were positive in this case of vCJD, but not in sCJD cases, including those with a heterozygous genotype (2). The Study We injected 18 RIII mice with 10% wt/vol frozen central nervous system tissue, 0.02 mL intracerebrally and 0.1 mL intraperitoneally, from a 129MV patient with a clinical case of vCJD (1). The vCJD tissue samples were provided by the NHS National Prion Clinic, University College London (UCL) Hospitals (London, UK), and MRC Prion Unit at UCL and sourced through the MRC Edinburgh Brain Bank (Edinburgh, Scotland, UK). The Brain Bank has full ethics approval and consent for the use of tissue in research (East of Scotland Research Ethics Service, Ref 16/ES/0084) and works within the framework of the Human Tissue (Scotland) Act 2006. We conducted inoculation, clinical scoring, and neuropathological and biochemical analysis of the mice as previously described (3-5). Animal studies were conducted according to the regulations of the UK Home Office Animals (Scientific Procedures) Act 1986. The isolate from the brain of the 129MV patient transmitted successfully; clinical and neuropathological signs associated with prion disease appeared in the mice. We compared the mean incubation period, neuropathological signs, and biochemical analysis with archived records of UK 129MM vCJD central nervous system transmissions and UK BSE transmissions. Methods used for inoculation, clinical scoring, and neuropathological and biochemical analysis of the mice were described in previous publications (3-5).
Journal of Biological Chemistry, 2004
There are two common forms of prion protein (PrP) in humans, with either methionine or valine at position 129. This polymorphism is a powerful determinant of the genetic susceptibility of humans toward both sporadic and acquired forms of prion disease and restricts propagation of particular prion strains. Despite its key role, we have no information on the effect of this mutation on the structure, stability, folding, and dynamics of the cellular form of PrP (PrP C). Here, we show that the mutation has no measurable effect on the folding, dynamics, and stability of PrP C. Our data indicate that the 129M/V polymorphism does not affect prion propagation through its effect on PrP C ; rather, its influence is likely to be downstream in the disease mechanism. We infer that the M/V effect is mediated through the conformation or stability of disease-related PrP (PrP Sc) or intermediates or on the kinetics of their formation. The prion diseases are a group of fatal neurodegenerative diseases that include scrapie in sheep and goats; bovine spongiform encephalopathy (BSE) 1 in cattle; and Creutzfeldt-Jakob disease (CJD), Gerstmann-Strä ussler-Scheinker disease, fatal familial insomnia (FFI), and kuru in humans. The human diseases may be inherited, arise sporadically, or be acquired through exposure to infectious prions (1, 2). Although rare in humans, intense interest has focused on these diseases both because of their unique biology and because of the occurrence of variant CJD, a new form of human prion disease, and the experimental evidence that it is caused by a BSE-like prion strain (3-5). According to the "protein-only" hypothesis (6), prions are composed principally or entirely of abnormal isoforms of hostencoded prion protein (PrP) (7). The disease-related isoform, PrP Sc , is derived from its normal cellular precursor, PrP C , by a
Histology and histopathology, 2007
Transmissible spongiform encephalopathies are a group of neural degenerative diseases that may be infectious, sporadic, or hereditary and are associated with an abnormally folded prion protein. Unfortunately at the current time it is not at all clear what the normal structure of the prion protein actually is or how it is toxic to cells. Extensive research on prion diseases has led to a dramatic increase in understanding of the pathogenesis of prion disorders, which will hopefully lead to the development of effective treatments. The inability to detect the disease in blood using current technology has made screening difficult. While fortunately there has been a decline in the number of clinical cases of transmissible variant CJD, evidence indicates that very long incubation periods can occur in humans so there may be a long slow, gradual epidemic. In particular, clinical cases in genotypes other than those homozygous for methionine at codon 129 of PRNP have not yet occurred, but such...
Human Mutation, 2000
Inherited prion diseases are characterized by mutations in the PRNP gene encoding the prion protein (PrP). As the other sporadic or infectious prion disease forms, they are almost all characterized by the accumulation in the brain of an abnormal misfolded form of the patient's PrP. Brain extracts can often transmit the disease once inoculated in a recipient animal. Inherited prion diseases with Creutzfeldt-Jakob disease (CJD) phenotype are autosomal forms, although sporadic cases have been reported. We report three novel mutations of the PRNP gene in unrelated patients with clinical and histopathologic features of CJD. The three mutations were missense: c635G>A (E196K), c656G>A (V203I) and c680G>C (E211Q). Familial history of neurologic disorders was evidenced for patients carrying the E196K and E211Q mutations. E196K would be predicted to have more severe effects on protein stability than V203I and E211Q. These mutations expand the spectrum of mutations in PRNP and reduce the proportion of CJD patients in whom genetic alterations have not been found.
Fatal Prion Disease in a Mouse Model of Genetic E200K Creutzfeldt-Jakob Disease
PLOS Pathogens, 2011
Genetic prion diseases are late onset fatal neurodegenerative disorders linked to pathogenic mutations in the prion protein-encoding gene, PRNP. The most prevalent of these is the substitution of Glutamate for Lysine at codon 200 (E200K), causing genetic Creutzfeldt-Jakob disease (gCJD) in several clusters, including Jews of Libyan origin. Investigating the pathogenesis of genetic CJD, as well as developing prophylactic treatments
Comparative Study of Prions in Iatrogenic and Sporadic Creutzfeldt-Jakob Disease
Journal of clinical & cellular immunology, 2014
Differentiating iatrogenic Creutzfeldt-Jakob disease (iCJD) from sporadic CJD (sCJD) would be useful for the identification and prevention of human-to-human prion transmission. Currently, the diagnosis of iCJD depends on identification of a recognized source of contamination to which patients have been exposed, in addition to fulfilling basic requirements for the establishment of diagnosis of CJD. Attempts to identify differences in clinical manifestations, neuropathological changes and pathological prion protein (PrP(Sc)) between iCJD and sCJD have been unsuccessful. In the present study, using a variety of more sophisticated methods including sucrose step gradient sedimentation, conformational stability immunoassay, protein misfolding cyclic amplification (PMCA), fragment-mapping, and transmission study, we show no significant differences in gel profiles, oligomeric state, conformational stability and infectivity of PrP(Sc) between iCJD and sCJD. However, using PMCA, we find that ...